Fig. 2: MEN2 patients’ functional kinome demonstrates pathogenic variant specificity.

a Unsupervised heatmap of phospho-site targets identified within serine/threonine and tyrosine PamGene peptide arrays of MEN2 patient derived RET probed PBMCs. Signal intensities are represented as Z-scores calculated from the mean of all phosphorylation scores within the data set. b Volcano plots of phospho-sites with altered phosphorylation states when analysed between (left) MEN2A and controls (ctrls), (middle) MEN2B and ctrls, and (right) MEN2A against MEN2B. Significantly downregulated phospho-sites are shown in red. Significantly upregulated phospho-sites are shown in teal. p < 0.05. c, d Venn diagrams showing the number, percentage and overlap of significantly altered phospho-sites for each (c) disease and (d) pathogenic variant, all groups were compared to ctrls. e–h Phospho-sites with the greatest change in phosphorylation across all MEN2 patients are depicted against ctrls. Each point represents an individual patient. Colour represents pathogenic variant (C609x-C634x variants are shown in green, S891A in red, V804M in light blue and M918T in dark blue). * p < 0.05, ** p < 0.005, *** p < 0.0005, analysed by an unpaired t test with Welch’s correction. Locations of Tyrosine (Y, White circle), Serine (S, white triangle) and Threonine (T, black triangle) phosphorylation sites located within the phospho-peptides shown are depicted below each respective graph. i Charts show 2-fold change in differential activity for upstream kinases predicted upon PamGene results for each pathogenic variant. Kinases with the 10% highest and 10% lowest change in predicted activity are described below each plot.